U.S. patent application number 17/366512 was filed with the patent office on 2022-06-02 for inhibitory immune receptor inhibition methods and compositions.
The applicant listed for this patent is The Board of Trustees of the Leland Stanford Junior University. Invention is credited to Carolyn R. Bertozzi, Elliot C. Woods, Han Xiao.
Application Number | 20220169724 17/366512 |
Document ID | / |
Family ID | 1000006140426 |
Filed Date | 2022-06-02 |
United States Patent
Application |
20220169724 |
Kind Code |
A1 |
Woods; Elliot C. ; et
al. |
June 2, 2022 |
Inhibitory Immune Receptor Inhibition Methods and Compositions
Abstract
Provided are methods relating to the inhibition of inhibitory
immune receptors. Aspects of the present disclosure include methods
that include administering to an individual receiving an antibody
therapy an inhibitory immune receptor inhibitor. Also provided are
compositions and kits that find use, e.g., in practicing the
methods of the present disclosure.
Inventors: |
Woods; Elliot C.;
(Burlingame, CA) ; Xiao; Han; (East Palo Alto,
CA) ; Bertozzi; Carolyn R.; (Stanford, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
The Board of Trustees of the Leland Stanford Junior
University |
Stanford |
CA |
US |
|
|
Family ID: |
1000006140426 |
Appl. No.: |
17/366512 |
Filed: |
July 2, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16307428 |
Dec 5, 2018 |
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PCT/US2017/040483 |
Jun 30, 2017 |
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17366512 |
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62357653 |
Jul 1, 2016 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07K 16/2851 20130101;
C07K 16/2887 20130101; G01N 2333/70503 20130101; A61P 35/00
20180101; C07K 2317/24 20130101; C07K 2317/732 20130101; C07K
2317/76 20130101; C07K 2317/73 20130101; C07K 16/32 20130101; G01N
33/57492 20130101; C07K 16/2803 20130101; A61K 2039/505 20130101;
A61K 2039/507 20130101 |
International
Class: |
C07K 16/28 20060101
C07K016/28; A61P 35/00 20060101 A61P035/00; C07K 16/32 20060101
C07K016/32; G01N 33/574 20060101 G01N033/574 |
Goverment Interests
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH
[0002] This invention was made with Government support under
contract GM059907 awarded by the National Institutes of Health. The
Government has certain rights in the invention.
Claims
1.-60. (canceled)
61. A method of treating cancer in an individual having cancer and
receiving an antibody therapy for the cancer, the method comprising
administering to the individual a therapeutically effective amount
of an inhibitory immune receptor inhibitor, wherein the inhibitory
immune receptor inhibitor inhibits Siglec-7 or Siglec-9.
62. The method according to claim 61, wherein the inhibitory immune
receptor inhibitor is an antibody.
63. The method according to claim 62, wherein the antibody is
selected from the group consisting of: an IgG, a single chain Fv
(scFv), Fab, F(ab').sub.2, or (scFv').sub.2.
64. The method according to claim 63, wherein the antibody is an
IgG.
65. The method according to claim 61, wherein the inhibitory immune
receptor inhibitor inhibits Siglec-7.
66. The method according to claim 61, wherein the inhibitory immune
receptor inhibitor inhibits Siglec-9.
67. The method according to claim 61, wherein the individual has
breast cancer, ovarian cancer, gastric cancer, colon cancer, or
renal carcinoma.
68. The method according to claim 61, wherein the individual is
receiving an antibody therapy that comprises administering to the
individual an antibody that specifically binds to a
tumor-associated antigen or a tumor-specific antigen.
69. The method according to claim 61, wherein the individual is
receiving an antibody therapy that comprises administering to the
individual an antibody that specifically binds an antigen selected
from the group consisting of: human epidermal growth factor
receptor 2 (HER2), CD19, CD22, CD30, CD33, CD56, CD66/CEACAM5,
CD70, CD74, CD79b, CD138, Nectin-4, Mesothelin, Transmembrane
glycoprotein NMB (GPNMB), Prostate-Specific Membrane Antigen
(PSMA), SLC44A4, CA6, CA-IX, an integrin, C-X-C chemokine receptor
type 4 (CXCR4), and neuropilin-1 (NRP1).
70. The method according to claim 61, wherein the inhibitory immune
receptor inhibitor inhibits Siglec-7, and wherein the method
comprises determining the abundance of Siglec-7 ligands present on
cells targeted by the antibody therapy prior to administering the
inhibitory immune receptor inhibitor.
71. The method according to claim 70, wherein the abundance of
Siglec-7 ligands is determined on a biopsy sample.
72. The method according to claim 71, wherein determining the
abundance of Siglec-7 ligands comprises incubating cells of the
biopsy sample with a reagent comprising an extracellular domain of
Siglec-7.
73. The method according to claim 61, wherein the inhibitory immune
receptor inhibitor inhibits Siglec-9, and wherein the method
comprises determining the abundance of Siglec-9 ligands present on
cells targeted by the antibody therapy prior to administering the
inhibitory immune receptor inhibitor.
74. The method according to claim 73, wherein the abundance of
Siglec-9 ligands is determined on a biopsy sample.
75. The method according to claim 74, wherein determining the
abundance of Siglec-9 ligands comprises incubating cells of the
biopsy sample with a reagent comprising an extracellular domain of
Siglec-9.
76. A pharmaceutical composition, comprising: an inhibitory immune
receptor inhibitor, wherein the inhibitory immune receptor
inhibitor inhibits Siglec-7 or Siglec-9; a therapeutic antibody;
and a pharmaceutically acceptable carrier.
77. The pharmaceutical composition of claim 76, wherein the
inhibitory immune receptor inhibitor is an antibody selected from
the group consisting of: an IgG, a single chain Fv (scFv), Fab,
F(ab')2, or (scFv')2.
78. The pharmaceutical composition of claim 76, wherein the
inhibitory immune receptor inhibitor inhibits Siglec-7.
79. The pharmaceutical composition of claim 76, wherein the
inhibitory immune receptor inhibitor inhibits Siglec-9.
80. A kit, comprising: a pharmaceutical composition comprising an
inhibitory immune receptor inhibitor, wherein the inhibitory immune
receptor inhibitor inhibits Siglec-7 or Siglec-9; and instructions
for using the composition in combination with an antibody therapy
being administered to an individual.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Patent Application No. 62/357,653 filed Jul. 1, 2016, which
application is incorporated herein by reference in its
entirety.
INTRODUCTION
[0003] Therapies that enhance the immune response to cancer are
proving revolutionary in the fight against intractable tumors.
Immune cells integrate signals from activating and inhibitory
receptors to determine their response to a challenging
target--activating signals alert them to the presence of pathology
while inhibitory signals tell the cell that it has confronted a
healthy "self". Successful tumors evolve mechanisms to thwart
immune cell recognition, often by overexpressing ligands for
inhibitory receptors. This discovery has led to new therapeutic
strategies aimed at blocking inhibitory immune cell signaling, as
embodied in clinically approved T cell checkpoint inhibitors
targeting PD-1 and CTLA-4. Ongoing pre-clinical studies have
focused on combining therapies targeting multiple immunologic
pathways. For example, antibodies against PD-1/PD-L1 in combination
with those targeting other T cell checkpoint inhibitors demonstrate
improved anti-tumor activity in syngeneic tumor models. A
complement to these interventions are therapies targeting innate
immune cells, particularly natural killer (NK) cells, macrophages
and dendritic cells.
SUMMARY
[0004] Provided are methods relating to the inhibition of
inhibitory immune receptors. Aspects of the present disclosure
include methods that include administering to an individual
receiving an antibody therapy an inhibitory immune receptor
inhibitor. Also provided are compositions and kits that find use,
e.g., in practicing the methods of the present disclosure.
BRIEF DESCRIPTION OF THE FIGURES
[0005] FIG. 1 shows data demonstrating the potentiation of
trastuzumab-dependent cytotoxicity in BT-20 cells by treatment with
Siglec blocking antibodies.
[0006] FIG. 2 shows data demonstrating the potentiation of
rituximab-dependent cytotoxicity in Ramos cells by treatment with
Siglec blocking antibodies.
[0007] FIG. 3 shows flow cytometry data demonstrating that BT-20
cells are rich in Siglec-7 and Siglec-9 ligands.
[0008] FIG. 4 shows flow cytometry data demonstrating that Ramos
cells are rich in Siglec-7 and Siglec-9 ligands.
[0009] FIG. 5 shows data indicating that Siglec-7 ligand abundance
predicts an increase in ADCC by a trastuzumab-sialidase conjugate.
HER2 expression level is indicated by number of `+`s. ADCC
potentiation is most pronounced for HER2-low cell line, e.g., as
seen in comparing MDA-MB-231 cells to SKBR3 cells.
DETAILED DESCRIPTION
[0010] Provided are methods relating to the inhibition of
inhibitory immune receptors. Aspects of the present disclosure
include methods that include administering to an individual
receiving an antibody therapy an inhibitory immune receptor
inhibitor. Also provided are compositions and kits that find use,
e.g., in practicing the methods of the present disclosure.
[0011] Before the methods, compositions and kits of the present
disclosure are described in greater detail, it is to be understood
that the methods, compositions and kits are not limited to
particular embodiments described, as such may, of course, vary. It
is also to be understood that the terminology used herein is for
the purpose of describing particular embodiments only, and is not
intended to be limiting, since the scope of the methods,
compositions and kits will be limited only by the appended
claims.
[0012] Where a range of values is provided, it is understood that
each intervening value, to the tenth of the unit of the lower limit
unless the context clearly dictates otherwise, between the upper
and lower limit of that range and any other stated or intervening
value in that stated range, is encompassed within the methods,
compositions and kits. The upper and lower limits of these smaller
ranges may independently be included in the smaller ranges and are
also encompassed within the methods, compositions and kits, subject
to any specifically excluded limit in the stated range. Where the
stated range includes one or both of the limits, ranges excluding
either or both of those included limits are also included in the
methods, compositions and kits.
[0013] Certain ranges are presented herein with numerical values
being preceded by the term "about." The term "about" is used herein
to provide literal support for the exact number that it precedes,
as well as a number that is near to or approximately the number
that the term precedes. In determining whether a number is near to
or approximately a specifically recited number, the near or
approximating unrecited number may be a number which, in the
context in which it is presented, provides the substantial
equivalent of the specifically recited number.
[0014] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which the methods, compositions and
kits belong. Although any methods, compositions and kits similar or
equivalent to those described herein can also be used in the
practice or testing of the methods, compositions and kits,
representative illustrative methods, compositions and kits are now
described.
[0015] All publications and patents cited in this specification are
herein incorporated by reference as if each individual publication
or patent were specifically and individually indicated to be
incorporated by reference and are incorporated herein by reference
to disclose and describe the materials and/or methods in connection
with which the publications are cited. The citation of any
publication is for its disclosure prior to the filing date and
should not be construed as an admission that the present methods,
compositions and kits are not entitled to antedate such
publication, as the date of publication provided may be different
from the actual publication date which may need to be independently
confirmed.
[0016] It is noted that, as used herein and in the appended claims,
the singular forms "a", "an", and "the" include plural referents
unless the context clearly dictates otherwise. It is further noted
that the claims may be drafted to exclude any optional element. As
such, this statement is intended to serve as antecedent basis for
use of such exclusive terminology as "solely," "only" and the like
in connection with the recitation of claim elements, or use of a
"negative" limitation.
[0017] It is appreciated that certain features of the methods,
compositions and kits, which are, for clarity, described in the
context of separate embodiments, may also be provided in
combination in a single embodiment. Conversely, various features of
the methods, compositions and kits, which are, for brevity,
described in the context of a single embodiment, may also be
provided separately or in any suitable sub-combination. All
combinations of the embodiments are specifically embraced by the
present disclosure and are disclosed herein just as if each and
every combination was individually and explicitly disclosed, to the
extent that such combinations embrace operable processes and/or
compositions. In addition, all sub-combinations listed in the
embodiments describing such variables are also specifically
embraced by the present methods, compositions and kits and are
disclosed herein just as if each and every such sub-combination was
individually and explicitly disclosed herein.
[0018] As will be apparent to those of skill in the art upon
reading this disclosure, each of the individual embodiments
described and illustrated herein has discrete components and
features which may be readily separated from or combined with the
features of any of the other several embodiments without departing
from the scope or spirit of the present methods. Any recited method
can be carried out in the order of events recited or in any other
order that is logically possible.
Methods
[0019] As summarized above, methods relating to inhibitory immune
receptor inhibition are provided. In certain aspects, the methods
include administering to an individual receiving an antibody
therapy an inhibitory immune receptor inhibitor. Details of such
methods will now be described.
[0020] As used herein, an "inhibitory immune receptor" is a
receptor present on an immune cell that negatively regulates an
immune response. According to certain embodiments, the inhibitory
immune receptor inhibitor inhibits an inhibitory immune receptor
present on an immune cell selected from a natural killer (NK) cell,
a macrophage, a monocyte, a neutrophil, a dendritic cell, a T cell,
a B cell, a mast cell, a basophil, and an eosinophil.
[0021] Examples of inhibitory immune receptors which may be
inhibited according to the methods of the present disclosure
include inhibitory immune receptors of the Ig superfamily,
including but not limited to: CD200R, CD300a (IRp60; mouse MAIR-I),
CD300f (IREM-1), CEACAM1 (CD66a), Fc.gamma.RIIb, ILT-2 (LIR-1;
LILRB1; CD85j), ILT-3 (LIR-5; CD85k; LILRB4), ILT-4 (LIR-2;
LILRB2), ILT-5 (LIR-3; LILRB3; mouse PIR-B); LAIR-1, PECAM-1
(CD31), PILR-.alpha. (FDF03), SIRL-1, and SIRP-.alpha.. Further
examples of inhibitory immune receptors which may be inhibited
according to the methods of the present disclosure include sialic
acid-binding Ig-like lectin (Siglec) receptors, including but not
limited to: Siglec-2, Siglec-3 (CD33), Siglec-5, mouse Siglec-f,
Siglec-6, Siglec-7, Siglec-8, Siglec-9, mouse Siglec-e, Siglec-10,
mouse Siglec-g, Siglec-11, and Siglec-12. Additional examples of
inhibitory immune receptors which may be inhibited according to the
methods of the present disclosure include C-type lectins, including
but not limited to: CLEC4A (DCIR), Ly49Q and MICL. Details
regarding inhibitory immune receptors may be found, e.g., in
Steevels et al. (2011) Eur. J. Immunol. 41(3):575-587.
[0022] In some embodiments, the inhibitory immune receptor is a
receptor for which the ligand is selected from an oligosaccharide,
a polysaccharide (or "glycan", that is, a molecule containing
monosaccharides linked glycosidically), a glycoprotein, a
glycolipid, and a ganglioside. In certain aspects, the inhibitory
immune receptor is a receptor for which the ligand has a terminal
sialic acid residue. According to certain embodiments, the
inhibitory immune receptor is a receptor for which the ligand is a
sialoglycan.
[0023] According to certain embodiments, the methods of the present
disclosure include administering two or more inhibitory immune
receptor inhibitors to the individual receiving an antibody
therapy. As just one example, the methods may include administering
to the individual a Siglec-7 inhibitor and a Siglec-9 inhibitor
(e.g., concurrently (e.g., as part of the same or different
compositions) or sequentially).
[0024] By inhibitory immune receptor "inhibitor" is meant an agent
that reduces or abolishes the biological activity of an inhibitory
immune receptor. The inhibitor employed may vary depending upon the
nature of the inhibitory immune receptor. Non-limiting examples of
inhibitors that may be employed include small molecules, ligands,
and antibodies.
[0025] According to certain embodiments, the inhibitor is a small
molecule. As used herein, a "small molecule" is a compound having a
molecular weight of 1000 atomic mass units (amu) or less. In some
embodiments, the small molecule is 750 amu or less, 500 amu or
less, 400 amu or less, 300 amu or less, or 200 amu or less. In
certain aspects, the small molecule is not made of repeating
molecular units such as are present in a polymer.
[0026] In certain aspects, the inhibitor is an antibody. The terms
"antibody" and "immunoglobulin" include antibodies or
immunoglobulins of any isotype (e.g., IgG (e.g., IgG1, IgG2, IgG3
or IgG4), IgE, IgD, IgA, IgM, etc.), whole antibodies (e.g.,
antibodies composed of a tetramer which in turn is composed of two
dimers of a heavy and light chain polypeptide); single chain
antibodies; fragments of antibodies (e.g., fragments of whole or
single chain antibodies) which retain specific binding to the
inhibitory immune receptor, including, but not limited to single
chain Fv (scFv), Fab, F(ab').sub.2, (scFv').sub.2, and diabodies;
chimeric antibodies; monoclonal antibodies, human antibodies,
humanized antibodies (e.g., humanized whole antibodies, humanized
half antibodies, or humanized antibody fragments); and fusion
proteins comprising an antigen-binding portion of an antibody and a
non-antibody protein. The antibodies may be detectably labeled,
e.g., with an in vivo imaging agent, a radioisotope, an enzyme
which generates a detectable product, a fluorescent protein, and
the like. The antibodies may be further conjugated to other
moieties, such as members of specific binding pairs, e.g., biotin
(member of biotin-avidin specific binding pair), and the like.
[0027] In certain aspects, the inhibitor (e.g., a small molecule,
an antibody, a sialic acid derivative, etc.) inhibits the
biological activity of an inhibitory immune receptor by binding
(e.g., specifically binding) to the inhibitory immune receptor. As
used herein, an inhibitor that "specifically binds" the inhibitory
immune receptor or is "specific" for the inhibitory immune receptor
refers to an inhibitor that binds the inhibitory immune receptor
with greater affinity than with other receptors. According to
certain embodiments, the inhibitor exhibits a binding affinity to
the inhibitory immune receptor of a K.sub.d of less than or equal
to about 10.sup.-5 M, less than or equal to about 10.sup.-6 M, or
less than or equal to about 10.sup.-7 M, or less than or equal to
about 10.sup.-8 M, or less than or equal to about 10.sup.-9 M,
10.sup.-10 M, 10.sup.-11 M, or 10.sup.-12 M or less. Such
affinities may be readily determined using conventional techniques,
such as by equilibrium dialysis, surface plasmon resonance (SPR)
technology (e.g., using the BIAcore 2000 instrument, using general
procedures outlined by the manufacturer), radioimmunoassay, or by
another method.
[0028] In some embodiments, the inhibitor may be a known inhibitor
of the inhibitory immune receptor of interest. In other
embodiments, the inhibitor is identified, e.g., using a suitable
approach for screening small molecules (e.g., by screening a
combinatorial library of small molecules), antibodies (e.g., by
phage or yeast display of antibody libraries), ligands, or the like
for the ability to inhibit (e.g., by binding) the inhibitory immune
receptor. The readout for such screening approaches will vary
depending upon the inhibitory immune receptor of interest.
[0029] In certain aspects, the methods include administering a
sialic acid-binding Ig-like lectin (Siglec) inhibitor to the
individual receiving an antibody therapy. The Siglec inhibitor may
be an inhibitor of, e.g., any of Siglecs 1-17. According to certain
embodiments, the inhibitor inhibits Siglec-7 (UniProtKB-Q9Y286),
Siglec-9 (UniProtKB-Q9Y336), or both. Such an inhibitor may be an
antibody, a small molecule, a sialic acid derivative, or the like.
Siglec inhibitors are described, e.g., in Cagnoni et al. (2016)
Front. Oncol. 6:109, and include, e.g., Oxamido-Neu5Ac, BPC-Neu5Ac,
BPC-Neu5Ac-Dox liposome, 9-BPC-4-mNPC-Neu5Ac, and the like.
Rational structure-based Siglec-7 inhibitor design is described,
e.g., in Attrill et al. (2006) Biochem. J. 397(2):271-8.
High-affinity Siglec-7 inhibitors are described, e.g., in Prescher
et al. (2017) J. Med. Chem. 60(3):941-956.
[0030] In some embodiments, the inhibitory immune receptor
inhibitor is an antibody that inhibits Siglec-7, Siglec-9, or both.
Non-limiting examples of available Siglec-7 and Siglec-9 blocking
antibodies are provided in the Experimental section below. In
certain aspects, the administered antibody that inhibits Siglec-7,
Siglec-9, or both, is a polyclonal, monoclonal, humanized, fully
human, asymmetric, or heteromeric antibody, or an antibody having
any combination of such features to the extent possible. In some
embodiments, the antibody that inhibits Siglec-7, Siglec-9, or
both, is a whole antibody (e.g., an antibody composed of a tetramer
which in turn is composed of two dimers of a heavy and light chain
polypeptide), such as a whole IgG (e.g., IgG1, IgG2, IgG3 or IgG4),
IgE, IgD, IgA, IgM, etc. antibody. In other aspects, the antibody
that inhibits Siglec-7, Siglec-9, or both, is an antibody fragment,
non-limiting examples of which are single chain Fv (scFv), Fab,
F(ab').sub.2, (scFv').sub.2, and the like. The antibody that
inhibits Siglec-7, Siglec-9, or both, may be a known antibody. In
certain aspects, such an antibody is identified, e.g., using a
suitable approach for screening antibodies (e.g., by phage or yeast
display of antibody libraries), for the ability to bind Siglec-7,
Siglec-9, or both.
[0031] Antibodies that specifically bind an inhibitory immune
receptor of interest (e.g., Siglec-7, Siglec-9, or both) can be
prepared using a wide variety of techniques known in the art
including the use of hybridoma, recombinant, phage display
technologies, or a combination thereof. For example, an antibody
may be made and identified/produced using methods of phage display.
Phage display is used for the high-throughput screening of protein
interactions. Phages may be utilized to display antigen-binding
domains expressed from a repertoire or combinatorial antibody
library (e.g., human or murine). Phage expressing an antigen
binding domain that binds Siglec-7, Siglec-9, or both, can be
selected or identified with Siglec-7 and/or Siglec-9, e.g., using
labeled Siglec-7 and/or Siglec-9 bound or captured to a solid
surface or bead. Phage used in these methods are typically
filamentous phage including fd and M13 binding domains expressed
from phage with Fab, Fv (individual Fv region from light or heavy
chains) or disulfide stabilized Fv antibody domains recombinantly
fused to either the phage gene III or gene VIII protein. Exemplary
methods are set forth, for example, in U.S. Pat. No. 5,969,108,
Hoogenboom, H. R. and Chames, Immunol. Today 2000, 21:371; Nagy et
al. Nat. Med. 2002, 8:801; Huie et al., Proc. Natl. Acad. Sci. USA
2001, 98:2682; Lui et al., J. Mol. Biol. 2002, 315:1063, each of
which is incorporated herein by reference. Several publications
(e.g., Marks et al., Bio/Technology 1992, 10:779-783) have
described the production of high affinity human antibodies by chain
shuffling, as well as combinatorial infection and in vivo
recombination as a strategy for constructing large phage libraries.
In another embodiment, ribosomal display can be used to replace
bacteriophage as the display platform (see, e.g., Hanes et al.,
Nat. Biotechnol. 2000, 18:1287; Wilson et al., Proc. Natl. Acad.
Sci. USA 2001, 98:3750; or Irving et al., J. Immunol. Methods 2001,
248:31). Cell surface libraries may be screened for antibodies
(Boder et al., Proc. Natl. Acad. Sci. USA 2000, 97:10701; Daugherty
et al., J. Immunol. Methods 2000, 243:211). Such procedures provide
alternatives to traditional hybridoma techniques for the isolation
and subsequent cloning of monoclonal antibodies.
[0032] After phage selection, the antibody coding regions from the
phage can be isolated and used to generate whole antibodies,
including human antibodies, or any desired antigen binding
fragment, and expressed in any desired host, including mammalian
cells, insect cells, plant cells, yeast, and bacteria. For example,
techniques to recombinantly produce Fv, scFv, Fab, F(ab').sub.2,
and Fab' fragments may be employed using methods known in the
art.
[0033] By "antibody therapy" is meant that an antibody (which is
not an inhibitory immune receptor inhibitor) will be, has been,
and/or is being administered to the individual for a therapeutic
purpose. The antibody therapy will vary depending upon the
condition of the individual being treated. In some embodiments, the
antibody therapy includes the administration of an antibody (e.g.,
an IgG1, IgG2, IgG3, or IgG4 antibody) that specifically binds to
an antigen (e.g., a cell surface antigen, such as a protein or
non-protein cell surface antigen) on the surface of a cell relevant
to the medical condition of the individual. For example, the
antibody administered as part of the antibody therapy may bind to
an antigen present on the surface of a cell that contributes to the
medical condition, where binding of the antibody to the antigen
reduces or abolishes the cell's contribution to the medical
condition. According to certain embodiments, the antibody therapy
includes administering to the individual an antibody selected from
trastuzumab, cetuximab, daratumumab, girentuximab, panitumumab,
ofatumumab, and rituximab.
[0034] In certain aspects, the individual is receiving an antibody
therapy that includes administering to the individual an antibody
that induces antibody-dependent cellular cytotoxicity (ADCC). ADCC
is the killing of an antibody-coated target cell by a cytotoxic
effector cell (e.g., via a nonphagocytic process), characterized by
the release of the content of cytotoxic granules and/or by the
expression of cell death-inducing molecules. ADCC may be triggered
through interaction of target-bound antibodies (e.g., IgG (e.g.,
IgG1, IgG2, IgG3, or IgG4), IgA, or IgE antibodies) with certain Fc
receptors (FcRs), glycoproteins present on the effector cell
surface that bind the Fc region of immunoglobulins (Ig). Effector
cells that mediate ADCC include natural killer (NK) cells,
monocytes, macrophages, neutrophils, eosinophils and dendritic
cells. ADCC is a rapid effector mechanism whose efficacy is
dependent on a number of parameters (density and stability of the
antigen on the surface of the target cell; antibody affinity and
FcR-binding affinity). ADCC involving human IgG1, the most used IgG
subclass for therapeutic antibodies, has been shown to be dependent
on the glycosylation profile of its Fc portion and on the
polymorphism of Fc.gamma. receptors.
[0035] Non-limiting examples of antibodies that may be administered
to the individual as part of the antibody therapy include
Adecatumumab, Ascrinvacumab, Cixutumumab, Conatumumab, Daratumumab,
Drozitumab, Duligotumab, Durvalumab, Dusigitumab, Enfortumab,
Enoticumab, Figitumumab, Ganitumab, Glembatumumab, Intetumumab,
Ipilimumab, Iratumumab, Icrucumab, Lexatumumab, Lucatumumab,
Mapatumumab, Namatumab, Necitumumab, Nesvacumab, Ofatumumab,
Olaratumab, Panitumumab, Patritumab, Pritumumab, Radretumab,
Ramucirumab, Rilotumumab, Robatumumab, Seribantumab, Tarextumab,
Teprotumumab, Tovetumab, Vantictumab, Vesencumab, Votumumab,
Zalutumumab, Flanvotumab, Altumomab, Anatumomab, Arcitumomab,
Bectumomab, Blinatumomab, Detumomab, Ibritumomab, Minretumomab,
Mitumomab, Moxetumomab, Naptumomab, Nofetumomab, Pemtumomab,
Pintumomab, Racotumomab, Satumomab, Solitomab, Taplitumomab,
Tenatumomab, Tositumomab, Tremelimumab, Abagovomab, Igovomab,
Oregovomab, Capromab, Edrecolomab, Nacolomab, Amatuximab,
Bavituximab, Brentuximab, Cetuximab, Derlotuximab, Dinutuximab,
Ensituximab, Futuximab, Girentuximab, Indatuximab, Isatuximab,
Margetuximab, Rituximab, Siltuximab, Ublituximab, Ecromeximab,
Abituzumab, Alemtuzumab, Bevacizumab, Bivatuzumab, Brontictuzumab,
Cantuzumab, Cantuzumab, Citatuzumab, Clivatuzumab, Dacetuzumab,
Demcizumab, Dalotuzumab, Denintuzumab, Elotuzumab, Emactuzumab,
Emibetuzumab, Enoblituzumab, Etaracizumab, Farletuzumab,
Ficlatuzumab, Gemtuzumab, Imgatuzumab, Inotuzumab, Labetuzumab,
Lifastuzumab, Lintuzumab, Lorvotuzumab, Lumretuzumab, Matuzumab,
Milatuzumab, Nimotuzumab, Obinutuzumab, Ocaratuzumab, Otlertuzumab,
Onartuzumab, Oportuzumab, Parsatuzumab, Pertuzumab, Pinatuzumab,
Polatuzumab, Sibrotuzumab, Simtuzumab, Tacatuzumab, Tigatuzumab,
Trastuzumab, Tucotuzumab, Vandortuzumab, Vanucizumab, Veltuzumab,
Vorsetuzumab, Sofituzumab, Catumaxomab, Ertumaxomab, Depatuxizumab,
Ontuxizumab, Blontuvetmab, Tamtuvetmab, or an antigen-binding
variant thereof. As used herein, "variant" is meant the antibody
binds to the target/antigen (e.g., HER2 for trastuzumab) but has
fewer or more amino acids than the parental antibody, has one or
more amino acid substitutions relative to the parental antibody, or
a combination thereof.
[0036] In some embodiments, the individual is receiving an antibody
therapy that includes administering to the individual an antibody
set forth in Table 1 below approved for treating cancer, or an
antigen-binding variant thereof. Also provided in Table 1 is the
corresponding tumor-associated antigen or tumor-specific antigen to
which the therapeutic antibody specifically binds, as well as the
type of cancer for which the antibody is approved for
treatment.
TABLE-US-00001 TABLE 1 Antibodies approved for treating cancer
Antigen Cancer Types Antibody BCR-ABL CML Imatinib, Dasatinib ALL
Nilotinib, Bosutinib Ponatinib CD19 ALL Blinatumomab CD20 NHL, CLL
Rituximab B-cell NHL Ofatumumab pre-B ALL .sup.90Y-Ibritumomab
.sup.131I-Tositumomab CD30 Hodgkin's lymphoma Brentuximab vedotin
CD33 AML Gemtuzumab ozogamicin CD52 CLL Alemtuzumab CTLA-4
Unresectable or metastatic Ipilimumab melanoma EGFR CRC Cetuximab
Head and Neck Panitumumab EpCAM Malignant ascites Catumaxomab HER2
Breast Trastuzumab Pertuzumab PAP Prostate Sipuleucel-T PD-1
Metastatic melanoma Nivolumab NSCLC Pembrolizumab VEGF Breast,
Cervical Bevacizumab CRC, NSCLC RCC, Ovarian Glioblastoma VEGF-R2
Gastric Ramucirumab NSCLC
[0037] Abbreviations for Table 1 are as follows: ALL, acute
lymphoblastic leukemia; AML, acute myelogenous leukemia; BCR-ABL,
breakpoint cluster region Abelson tyrosine kinase; CLL, chronic
lymphocytic leukemia; CTLA-4, cytotoxic T-lymphocyte-associated
antigen 4; CRC, colorectal cancer; EGFR, epidermal growth factor
receptor; EpCAM, epithelial cell adhesion molecule; HER2, human
epidermal growth factor receptor 2; NHL, non-Hodgkin's lymphoma;
NSCLC, non-small cell lung cancer; PAP, prostatic acid phosphatase;
PD-1, programmed cell death receptor 1; RC, renal cell carcinoma;
VEGF, vascular endothelial growth factor; VEGF-R2, vascular
endothelial growth factor receptor 2.
[0038] In certain aspects, the individual is receiving an antibody
therapy that includes administering to the individual an antibody
set forth in Table 2 below or an antigen-binding variant thereof.
Also provided in Table 2 is the corresponding tumor-associated
antigen or tumor-specific antigen to which the therapeutic antibody
specifically binds, as well as an example cancer type which may be
treated using the antibody.
TABLE-US-00002 TABLE 2 Additional antibodies, cell surface
molecules, and cancer types Antigen Cancer types Antibody A2aR
NSCLC PBF-509 AKAP4 NSCLC Preclinical Ovarian BAGE Glioblastoma
Preclinical Ovarian BORIS Prostate, Lung Preclinical Esophageal
CD22 ALL Epratuzumab Moxetumomab Inotuzumab ozogamicin CD73
Advanced solid tumors MEDI9447 CD137 Advanced solid tumors Urelumab
PF-05082566 CEA CRC PANVAC .TM. Ad5-[E1-, E2b-]-CEA(6D) CS1
Multiple myeloma Elotuzumab CTLA-4 Malignant mesothelioma
Tremelimumab EBAG9 Bladder Preclinical EGF NSCLC CIMAvax EGFR NSCLC
Necitumumab GAGE Cervical Preclinical GD2 Neuroblastoma
Dinutuximab, hu3F8 Retinoblastorna hu14.18-IL-2, 3F8/OKT3BsAb
Melanoma other anti-GD2 CAR solid tumors GD2-KLH gp100 Melanoma
gp100:209-217(210M) HPV-16 Cervical HPV-16 (E6, E7) SCCHN TG4001,
Lm-LLO-E7 pNGVL4a-CRT/E7, INO-3112 HSP105 CRC Preclinical Bladder
IDH1 Glioma IDH1(R132H) p123-142 Idiotype NSCLC, Breast Racotumomab
(NeuGcGM3) Melanoma IDO1 Breast, Melanoma Indoximod NSCLC
INCB024360 IDO1 peptide vaccine KIR Lymphoma Lirilumab LAG-3
Breast, Hemato- BMS-986016 logical, Advanced IMP321 solid tumors
LY6K Gastric LY6K-177 peptide SCCHN LY6K, CDCA1, IMP3 MAGE-A3
Melanoma recMAGE-A3 NSCLC Zastumotide MAGE-C2 Gastric, Melanoma
Preclinical Multiple myeloma MAGE-D4 CRC Preclinical Melan-A
Melanoma MART-1 (26-35, 27L) MET NSCLC Onartuzumab Tivantinib MUC1
NSCLC, Breast Tecemotide, TG4010 Prostate PANVAC .TM. MUC4
Pancreatic Preclinical MUC-16 Ovarian Abagovomab Oregovomab
NY-ESO-1 Ovarian NY-ESO-1/ISCOMATRIX .TM. Melanoma rV-NY-ESO-1;
rF-NY-ESO-1 PD-1 B-cell lymphoma Pidiltzumab Melanoma, CRC AMP-224,
AMP-514 PD-L1 NSCLC, RCC BMS-936559, Atezoilzumab Bladder, Breast
Durvalumab, Avelumab Melanoma, SCCHN PRAME NSCLC Preclinical PSA
Prostate PROSTVAC .RTM.-VF ROR1 CLL, Pancreatic Preclinical Lung,
Breast Sialyl-Tn Breast Theratope SPAG-9 Prostate, CRC Preclinical
NSCLC, Ovarian SSX1 Prostate Preclinical Multiple myeloma Survivin
Melanoma EMD640744 Glioma, Solid tumors Trivalent peptide vaccine
Tripeptide vaccine Telomerase Pancreatic Tertomotide TIM-3
Melanoma, NHL NSCLC Preclinical VISTA Melanoma, Bladder Preclinical
WTI Ovarian, Uterine, AML WT1 peptide vaccine Multiple myeloma
XAGE-1b Prostate DC-based tumor vaccine 5T4 RCC, CRC TroVax .RTM.
Prostate Naptumomab estatenatox
[0039] Abbreviations for Table 2 are as follows: A2aR, adenosine
A2a receptor; AKAP4, A kinase anchor protein 4; AML, acute
myelogenous leukemia; ALL, acute lymphoblastic leukemia; BAGE, B
melanoma antigen; BORIS, brother of the regulator of imprinted
sites; CEA, carcinoembryonic antigen; CLL, chronic lymphocytic
leukemia; CRC, colorectal cancer; CS1, CD2 subset 1; CTLA-4,
cytotoxic T-lymphocyte-associated antigen 4; EBAG9, estrogen
receptor binding site associated antigen 9; EGF, epidermal growth
factor; EGFR, epidermal growth factor receptor; NSCLC, non-small
cell lung cancer; GAGE, G antigen; GD2, disialoganglioside GD2;
gp100, glycoprotein 100; HPV-16, human papillomavirus 16; HSP105,
heat-shock protein 105; IDH1, isocitrate dehydrogenase type 1;
IDO1, indoleamine-2,3-dioxygenase 1; KIR, killer cell
immunoglobulin-like receptor; LAG-3, lymphocyte activation gene 3;
LY6K, lymphocyte antigen 6 complex K; MAGE-A3, melanoma antigen 3;
MAGE-C2, melanoma antigen C2; MAGE-D4, melanoma antigen D4;
Melan-A/MART-1, melanoma antigen recognized by T-cells 1; MET,
N-methyl-N'-nitroso-guanidine human osteosarcoma transforming gene;
MUC1, mucin 1; MUC4, mucin 4; MUC16, mucin 16; NHL, non-Hodgkin
lymphoma; NY-ESO-1, New York esophageal squamous cell carcinoma 1;
PD-1, programmed cell death receptor 1; PD-L1, programmed cell
death receptor ligand 1; PRAME, preferentially expressed antigen of
melanoma; PSA, prostate specific antigen; RCC, renal cell
carcinoma; ROR1, receptor tyrosine kinase orphan receptor 1; SCCHN,
squamous cell carcinoma of the head and neck; SPAG-9,
sperm-associated antigen 9; SSX1, synovial sarcoma X-chromosome
breakpoint 1; TIM-3, T-cell immunoglobulin domain and mucin
domain-3; VISTA, V-domain immunoglobulin-containing suppressor of
T-cell activation; WT1, Wilms' Tumor-1; XAGE-1b, X chromosome
antigen 1 b.
[0040] In some embodiments, the individual is receiving an antibody
therapy that includes administering to the individual an antibody
set forth in Table 3 below, or a variant thereof. Also provided in
Table 3 is the corresponding tumor-associated antigen or
tumor-specific antigen to which the therapeutic antibody
specifically binds.
TABLE-US-00003 TABLE 3 Additional antibodies and corresponding cell
surface molecules Antibody Antigen oregovomab CA125 girentuximab
CAIX obinutuzumab CD20 ofatumumab CD20 rituximab CD20 alemtuzumab
CD52 ipilimumab CTLA-4 tremelimumab CTLA-4 cetuximab EGFR
necitumumab EGFR panitumumab EGFR zalutumumab EGFR edrecolomab
EpCAM (17-1A) farletuzumab FR-alpha pertuzumab Her2 trastuzumab
Her2 rilotumumab HGF figitumumab IGF-1 ganitumab IGF1R durvalumab
IGG1K bavituximab Phosphatidylserine onartuzumab scatter factor
receptor kinase bevacizumab VEGF-A ramucirumab VEGFR2
[0041] In certain aspects, the individual is receiving an antibody
therapy that includes administering to the individual an antibody
selected from trastuzumab, cetuximab, daratumumab, girentuximab,
panitumumab, ofatumumab, rituximab, and variants thereof.
[0042] According to certain embodiments, the antibody administered
as part of the antibody therapy may be conjugated to an agent,
e.g., a therapeutic agent, a labeling agent (e.g., an in vivo
imaging agent), or the like. For example, the antibody may be part
of an antibody-drug conjugate (ADC). Drugs of interest include
agents capable of affecting the function of a cell/tissue to which
the conjugate binds via specific binding of the antibody portion of
the conjugate to an antigen on the surface of the cell/tissue. For
example, the agent may boost the function of the cell/tissue to
which the conjugate specifically binds. Alternatively, when the
function of the cell/tissue is pathological, an agent that reduces
the function of the cell/tissue may be employed. In certain
aspects, a conjugate includes an agent that reduces the function of
a target cell/tissue by inhibiting cell proliferation and/or
killing the cell/tissue. Such agents may vary and include
cytostatic agents and cytotoxic agents, e.g., an agent capable of
killing a target cell tissue with or without being internalized
into a target cell.
[0043] In certain aspects, the antibody administered as part of the
antibody therapy is conjugated to a drug selected from an enediyne,
a lexitropsin, a duocarmycin, a taxane, a puromycin, a dolastatin,
a maytansinoid, and a vinca alkaloid. In some embodiments, the
cytotoxic agent is paclitaxel, docetaxel, CC-1065, CPT-11 (SN-38),
topotecan, doxorubicin, morpholino-doxorubicin, rhizoxin,
cyanomorpholino-doxorubicin, dolastatin-10, echinomycin,
combretastatin, calicheamicin, maytansine, maytansine DM1,
maytansine DM4, DM-1, an auristatin or other dolastatin
derivatives, such as auristatin E or auristatin F, AEB (AEB-071),
AEVB (5-benzoylvaleric acid-AE ester), AEFP (antibody-endostatin
fusion protein), MMAE (monomethylauristatin E), MMAF
(monomethylauristatin F), pyrrolobenzodiazepines (PBDs),
eleutherobin, netropsin, or any combination thereof.
[0044] The type of individual receiving the antibody therapy may
vary. In certain aspects, the individual is a "mammal" or
"mammalian," where these terms are used broadly to describe
organisms which are within the class mammalia, including the orders
carnivore (e.g., dogs and cats), rodentia (e.g., mice, guinea pigs,
and rats), and primates (e.g., humans, chimpanzees, and monkeys).
In some embodiments, the individual is a human.
[0045] According to certain embodiments, the individual has cancer.
In certain aspects, the individual has a cancer set forth in Table
1 or Table 2 above. In some embodiments, the cancer is selected
from breast cancer, ovarian cancer, gastric cancer, colon cancer,
and renal carcinoma. When the individual has cancer, the antibody
therapy may include administering to the individual an antibody
that binds to an antigen present on a cancer cell of the
individual. In some embodiments, the antibody binds to an antigen
set forth in Table 1, Table 2, or Table 3 above. By "cancer cell"
is meant a cell exhibiting a neoplastic cellular phenotype, which
may be characterized by one or more of, for example, abnormal cell
growth, abnormal cellular proliferation, loss of density dependent
growth inhibition, anchorage-independent growth potential, ability
to promote tumor growth and/or development in an immunocompromised
non-human animal model, and/or any appropriate indicator of
cellular transformation. "Cancer cell" may be used interchangeably
herein with "tumor cell", "malignant cell" or "cancerous cell", and
encompasses cancer cells of a solid tumor, a semi-solid tumor, a
primary tumor, a metastatic tumor, and the like. In certain
aspects, the cancer cell is a carcinoma cell. According to certain
embodiments, the cancer cell is selected from a breast cancer cell,
an ovarian cancer cell, a gastric cancer cell, a colon cancer cell,
and a renal carcinoma cell.
[0046] In certain aspects, when the individual has cancer, the
antibody therapy includes administering to the individual an
antibody (e.g., an ADCC-inducing antibody) that binds to a
tumor-associated antigen or a tumor-specific antigen. By
"tumor-associated antigen" is meant an antigen expressed on
malignant cells with limited expression on cells of normal tissues,
an antigen expressed at much higher density on malignant versus
normal cells, or an antigen that is developmentally expressed. In
certain aspects, the antibody therapy includes administering to the
individual an antibody that binds to a tumor-associated antigen or
a tumor-specific antigen selected from HER2, CD19, CD22, CD30,
CD33, CD56, CD66/CEACAM5, CD70, CD74, CD79b, CD138, Nectin-4,
Mesothelin, Transmembrane glycoprotein NMB (GPNMB),
Prostate-Specific Membrane Antigen (PSMA), SLC44A4, CA6, CA-IX, an
integrin, C-X-C chemokine receptor type 4 (CXCR4), cytotoxic
T-lymphocyte-associated protein 4 (CTLA-4), neuropilin-1 (NRP1),
matriptase, or any other tumor-associated or tumor-specific
antigens of interest. In some embodiments, the antibody therapy
includes administering to the individual an antibody set forth in
any of Table 1, Table 2, or Table 3 above, or an antigen-binding
variant thereof.
[0047] According to certain embodiments, the methods of the present
disclosure include determining the abundance of one or more
inhibitory immune receptor ligands present on cells targeted by the
antibody therapy prior to administering the inhibitory immune
receptor inhibitor. Ligand abundance may be determined using any
suitable approach and may vary depending upon the type of ligand to
be detected.
[0048] In certain aspects, ligand abundance is determined in vivo
(that is, in the individual). Approaches for detecting molecules
(e.g., proteins) of interest in vivo are known and include, e.g.,
in vivo imaging. For example, when the identity of the ligand is
known, an agent (e.g., an antibody) labeled with (e.g., conjugated
to) an in vivo imaging agent may be administered to the individual,
followed by detection of the ligand (via a detectable label of the
imaging agent) to determine the location and abundance of the
ligand. Suitable in vivo imaging agents include, but are not
limited to, those that find use in in vivo imaging applications
such as near-infrared (NIR) imaging, single photon emission
computed tomography (SPECT), and/or the like. Details regarding
suitable in vivo imaging approaches may be found, e.g., in Tunnell,
J. In Vivo Clinical Imaging and Diagnosis ISBN: 9780071626835.
[0049] In some embodiments, the abundance of one or more inhibitory
immune receptor ligands present on cells targeted by the antibody
therapy is determined in vitro. For example, ligand abundance may
be determined on a biopsy sample, e.g., on target cells/tissue
removed from the individual. Any suitable in vitro approach to
determine the abundance of the one or more inhibitory immune
receptor ligands may be employed. Such approaches include, but are
not limited to, flow cytometry, enzyme-linked immunosorbent assays
(ELISA), immunofluorescence, immunohistochemistry, etc. In certain
aspects, an antibody that specifically binds the ligand is
employed. In other aspects, a detection reagent that includes all
or a portion of an extracellular domain of the inhibitory immune
receptor is employed. For example, all or a portion of an
extracellular domain of the inhibitory immune receptor may be part
of a soluble fusion protein. Such a fusion protein may include all
or a portion of an extracellular domain of the inhibitory immune
receptor fused to a protein (e.g., a fragment crystallizable (Fc)
antibody fragment, a protein tag, etc.) to which a secondary
detection reagent may bind. The secondary detection reagent may be
a labeled secondary antibody, such as an antibody labeled with a
fluorescent dye, a radioisotope, an enzyme which generates a
detectable product, or the like.
[0050] In certain aspects, the inhibitory immune receptor inhibitor
is administered to the individual only upon determining that the
abundance of the one or more inhibitory immune receptor ligands on
cells targeted by the antibody therapy exceeds a threshold
abundance level. The threshold abundance level may be based on one
or more criteria. For example, when the ligand is detected using a
labeled detection reagent (e.g., a fluorescently-labeled reagent),
the threshold abundance level may be based at least in part on the
signal (e.g., fluorescence) intensity as compared to a control.
Fluorescence intensity, for example, may be determined by flow
cytometry, immunofluorescence staining, or the like. Suitable
controls to which the signal intensity may be compared include,
e.g., the signal intensity from a different labeled reagent (e.g.,
a different fluorophore) relating to a different molecule, e.g.,
which is known to not significantly vary in abundance between
normal cells and the type(s) of cells being evaluated in the biopsy
sample. In another example, the signal intensity relating to the
ligand on the surface of abnormal cells from the individual may be
compared to the fluorescence intensity relating to the same ligand
on the surface of control cells, which control cells may be, e.g.,
counterpart normal cells from the individual, counterpart normal
cells from a different individual, cells from a cell line for which
the abundance of the ligand is stable and has been established, or
the like.
[0051] The threshold abundance level may be a ratio of the signal
intensity relating to the inhibitory immune receptor ligand from
cells in the biopsy sample to a signal intensity relating to a
control molecule from cells in the biopsy sample. In certain
aspects, the threshold abundance level may be a ratio (e.g., 1:1,
1.5:1, 2:1, 2.5:1, 3:1, etc.) of the signal intensity relating to
the inhibitory immune receptor ligand from cells in the biopsy
sample to a signal intensity relating to the inhibitory immune
receptor ligand from control cells.
[0052] According to certain embodiments, the inhibitory immune
receptor inhibitor is administered to the individual only upon
determining that the abundance of the antigen to which the antibody
of the therapy binds exceeds a threshold level. In some
embodiments, the inhibitory immune receptor inhibitor is
administered to the individual only upon determining that the
abundance of the antigen to which the antibody of the therapy binds
is less than a threshold level. For example, the present inventors
have found that--in some instances (e.g., for certain therapeutic
antibodies)--the benefits of administering the inhibitory immune
receptor inhibitor are greatest when the abundance level of the
antigen to which the antibody of the therapy binds is moderate or
low. One such example is provided in the Experimental section
below.
[0053] As summarized above, the inhibitory immune receptor
inhibitor is administered to an individual receiving an antibody
therapy. According to certain embodiments, the inhibitory immune
receptor inhibitor is administered to the individual prior to the
onset of the antibody therapy, concurrently with the antibody
therapy, or both.
[0054] According to certain embodiments, the antibody administered
to the individual as part of the antibody therapy and/or the
inhibitory immune receptor inhibitor are administered according to
a dosing regimen approved for individual use. In some embodiments,
the administration of the inhibitory immune receptor inhibitor
permits the antibody administered to the individual as part of the
antibody therapy to be administered according to a dosing regimen
that involves one or more lower and/or less frequent doses, and/or
a reduced number of cycles as compared with that utilized when the
antibody is administered without administration of the inhibitory
immune receptor inhibitor.
[0055] In certain aspects, one or more doses of the antibody
administered to the individual as part of the antibody therapy and
the inhibitory immune receptor inhibitor are administered at the
same time; in some such embodiments, such agents may be
administered present in the same pharmaceutical composition. In
some embodiments, however, the antibody administered to the
individual as part of the antibody therapy and the inhibitory
immune receptor inhibitor are administered to the individual in
different compositions and/or at different times. For example, the
antibody administered to the individual as part of the antibody
therapy may be administered prior to administration of the
inhibitory immune receptor inhibitor (e.g., in a particular cycle).
Alternatively, the inhibitory immune receptor inhibitor may be
administered prior to administration of the antibody of the
antibody therapy (e.g., in a particular cycle). The second agent to
be administered may be administered a period of time that starts at
least 1 hour, 3 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72
hours, or up to 5 days or more after the administration of the
first agent to be administered.
[0056] In one example, the inhibitory immune receptor inhibitor is
administered to the individual for a desirable period of time prior
to administration of the antibody that is part of the antibody
therapy. In certain aspects, such a regimen "pre-blocks" the
biology activity of the inhibitory immune receptor to potentiate
ADCC resulting from the subsequent administration of an antibody
that induces ADCC. Such a period of time separating a step of
administering the inhibitory immune receptor inhibitor from a step
of administering the antibody of the antibody therapy is of
sufficient length to permit inhibition of the target inhibitory
immune receptor, desirably so that ADCC mediated by the antibody of
the antibody therapy is increased.
[0057] In some embodiments, administration of one agent is
specifically timed relative to administration of another agent. For
example, in some embodiments, a first agent is administered so that
a particular effect is observed (or expected to be observed, for
example based on population studies showing a correlation between a
given dosing regimen and the particular effect of interest).
[0058] In certain aspects, desired relative dosing regimens for
agents administered in combination may be assessed or determined
empirically, for example using ex vivo, in vivo and/or in vitro
models; in some embodiments, such assessment or empirical
determination is made in vivo, in a patient population (e.g., so
that a correlation is established), or alternatively in a
particular individual of interest.
[0059] By way of example, the antibody of the antibody therapy may
be administered a period of time after administration of the
inhibitory immune receptor inhibitor. The period of time may be
selected to be correlated with inhibition of the inhibitory immune
receptor by the inhibitory immune receptor inhibitor. In certain
aspects, the relevant period of time permits (e.g., is correlated
with) inhibition of the inhibitory immune receptor to a level that
is 90% or less, 80% or less, 70% or less, 60% or less, 50% or less,
40% or less, 30% or less, 20% or less, or 10% or less than that
observed on the relevant immune cells (e.g., NK cells) prior to (or
at the moment of) the administration of the antibody of the
antibody therapy.
[0060] In some embodiments, the inhibitory immune receptor
inhibitor and the antibody of the antibody therapy are administered
according to an intermittent dosing regimen including at least two
cycles. Where two or more agents are administered in combination,
and each by such an intermittent, cycling, regimen, individual
doses of different agents may be interdigitated with one another.
In certain aspects, one or more doses of the second agent is
administered a period of time after a dose of the first agent. In
some embodiments, each dose of the second agent is administered a
period of time after a dose of the first agent. In certain aspects,
each dose of the first agent is followed after a period of time by
a dose of the second agent. In some embodiments, two or more doses
of the first agent are administered between at least one pair of
doses of the second agent; in certain aspects, two or more doses of
the second agent are administered between al least one pair of
doses of the first agent. In some embodiments, different doses of
the same agent are separated by a common interval of time; in some
embodiments, the interval of time between different doses of the
same agent varies. In certain aspects, different doses of the
different agents are separated from one another by a common
interval of time; in some embodiments, different doses of the
different agents are separated from one another by different
intervals of time.
[0061] One exemplary protocol for interdigitating two intermittent,
cycled dosing regimens (e.g., for potentiating cellular
cytotoxicity dependent upon the antibody of the antibody therapy),
may include: (a) a first dosing period during which a
therapeutically effective amount a first agent is administered to a
patient; (b) a first resting period; (c) a second dosing period
during which a therapeutically effective amount of a second agent
and, optionally, a third agent, is administered to the patient; and
(d) a second resting period. By "therapeutically effective amount"
is meant a dosage sufficient to produce a desired result, e.g., an
amount sufficient to effect beneficial or desired therapeutic
(including preventative) results, such as a reduction in a symptom
of a disease or disorder associated with the target cell or a
population thereof, as compared to a control. An effective amount
can be administered in one or more administrations.
[0062] In some embodiments, the first resting period and second
resting period may correspond to an identical number of hours or
days. Alternatively, in some embodiments, the first resting period
and second resting period are different, with either the first
resting period being longer than the second one or, vice versa. In
some embodiments, each of the resting periods corresponds to 120
hours, 96 hours, 72 hours, 48 hours, 24 hours, 12 hours, 6 hours,
30 hours, 1 hour, or less. In some embodiments, if the second
resting period is longer than the first resting period, it can be
defined as a number of days or weeks rather than hours (for
instance 1 day, 3 days, 5 days, 1 week, 2, weeks, 4 weeks or
more).
[0063] If the first resting period's length is determined by
existence or development of a particular biological or therapeutic
event (e.g., inhibition of the inhibitory immune receptor), then
the second resting period's length may be determined on the basis
of different factors, separately or in combination. Exemplary such
factors may include type and/or stage of a cancer against which an
anti-tumor antibody therapy is administered; identity and/or nature
of a targeted tumor antigen, identity and/or properties (e.g.,
pharmacokinetic properties) of the first agent, and/or one or more
features of the patient's response to therapy with the first agent.
In some embodiments, length of one or both resting periods may be
adjusted in light of pharmacokinetic properties (e.g., as assessed
via plasma concentration levels) of one or the other of the
administered agents. For example, a relevant resting period might
be deemed to be completed when plasma concentration of the relevant
agent is below about 1 .mu.g/ml, 0.1 .mu.g/ml, 0.01 .mu.g/ml or
0.001 .mu.g/ml, optionally upon evaluation or other consideration
of one or more features of the individual's response.
[0064] In certain aspects, the number of cycles for which a
particular agent is administered may be determined empirically.
Also, in some embodiments, the precise regimen followed (e.g.,
number of doses, spacing of doses (e.g., relative to each other or
to another event such as administration of another therapy), amount
of doses, etc.) may be different for one or more cycles as compared
with one or more other cycles.
[0065] The antibody that is administered as part of the antibody
therapy and the inhibitory immune receptor inhibitor may be
administered together or independently via any suitable route of
administration. Such agents may be administered via a route of
administration independently selected from oral, parenteral (e.g.,
by intravenous, intra-arterial, subcutaneous, intramuscular, or
epidural injection), topical, or nasal administration. According to
certain embodiments, antibody that is administered as part of the
antibody therapy and the inhibitory immune receptor inhibitor are
both administered parenterally, either concurrently (in the same
pharmaceutical composition or separate pharmaceutical compositions)
or sequentially.
[0066] In certain aspects, the methods include administering to the
individual a further therapeutic agent in addition to the antibody
that is administered as part of the antibody therapy and the
inhibitory immune receptor inhibitor. Such administration may
include concurrently administering the further therapeutic agent
and one or both of the antibody that is administered as part of the
antibody therapy and the inhibitory immune receptor inhibitor, or
administering the further therapeutic agent sequentially with
respect to one or both of the antibody that is administered as part
of the antibody therapy and the inhibitory immune receptor
inhibitor. In some embodiments, the individual has cancer, and the
further therapeutic agent is an anti-cancer agent. Anti-cancer
agents of interest include, but are not limited to, anti-cancer
antibodies (e.g., any of the antibodies set forth in Tables 1, 2,
and 3 above), small molecule anti-cancer agents, or the like.
[0067] In some embodiments, the further therapeutic agent is a
small molecule anti-cancer agent selected from abiraterone,
bendamustine, bexarotene, bortezomib, clofarabine, decitabine,
exemestane, temozolomide, afatinib, axitinib, bosutinib,
cabozantinib, crizotinib, dabrafenib, dasatinib, erlotinib,
gefitinib, ibrutinib, imatinib, lapatinib, nilotinib, pazopanib,
ponatinib, regorafenib, ruxolitinib, sorafenib, sunitinib,
vandetanib, vemurafenib, enzalutamide, fulvestrant, epirubicin,
ixabepilone, nelarabine, vismodegib, cabazitaxel, pemetrexed,
azacitidine, carfilzomib, everolimus, temsirolimus, eribulin,
omacetaxine, trametinib, lenalidomide, pomalidomide, romidepsin,
vorinostat, brigatinib, ribociclib, midostaurin, telotristat ethyl,
niraparib, cabozantinib, lenvatinib, rucaparib, granisetron,
dronabinol, venetoclax, alectinib, cobimetinib, panobinostat,
palbociclib, talimogene laherparepvec, lenvatinib, trifluridine and
tipiracil, ixazomib, sonidegib, osimertinib, rolapitant, uridine
triacetate, trabectedin, netupitant and palonosetron, belinostat,
ibrutinib, olaparib, idelalisib, and ceritinib.
[0068] In certain aspects, the further therapeutic agent is an
immune checkpoint inhibitor. Immune checkpoint inhibitors of
interest include, but are not limited to, inhibitors (e.g.,
antibodies) that target PD-1, PD-1, CTLA-4, TIM3, LAG3, or a member
of the B7 family.
[0069] According to certain embodiments, the antibody that is
administered as part of the antibody therapy, the inhibitory immune
receptor inhibitor, and a further therapeutic agent are
administered according to a dosing regimen approved for individual
use. In some embodiments, the administration of the further
therapeutic agent permits the antibody that is administered as part
of the antibody therapy, the inhibitory immune receptor inhibitor,
or both, administered to the individual to be administered
according to a dosing regimen that involves one or more lower
and/or less frequent doses, and/or a reduced number of cycles as
compared with that utilized when the antibody that is administered
as part of the antibody therapy, the inhibitory immune receptor
inhibitor, or both, is administered without administration of the
further therapeutic agent. In certain aspects, the administration
of the antibody that is administered as part of the antibody
therapy, the inhibitory immune receptor inhibitor, or both, permits
the further therapeutic agent administered to the individual to be
administered according to a dosing regimen that involves one or
more lower and/or less frequent doses, and/or a reduced number of
cycles as compared with that utilized when the further therapeutic
agent is administered without administration of the antibody that
is administered as part of the antibody therapy, the inhibitory
immune receptor inhibitor, or both.
Compositions
[0070] As summarized above, aspects of the present disclosure
include pharmaceutical compositions. According to certain
embodiments, a pharmaceutical composition of the present disclosure
includes an inhibitory immune receptor inhibitor, a therapeutic
antibody, and a pharmaceutically acceptable carrier. The
pharmaceutical compositions generally include a therapeutically
effective amount of the inhibitory immune receptor inhibitor and
the therapeutic antibody.
[0071] Any of the inhibitory immune receptor inhibitors described
above may be present in a pharmaceutical composition of the present
disclosure. For example, the inhibitory immune receptor inhibitor
may be any of the antibodies or small molecules described above.
When the inhibitory immune receptor inhibitor is an antibody, the
antibody may be an IgG (e.g., an IgG1, IgG2, IgG3 or IgG4
antibody), a single chain Fv (scFv), Fab, (Fab)2, (scFv')2, or the
like. The antibody may be a monoclonal antibody, a humanized
antibody, a human antibody, etc.
[0072] An inhibitory immune receptor inhibitor present in a
pharmaceutical composition of the present disclosure may
specifically bind to an inhibitory immune receptor present on an
immune cell selected from a natural killer (NK) cell, a macrophage,
a monocyte, a neutrophil, a dendritic cell, a T cell, a B cell, a
mast cell, a basophil, and an eosinophil. In certain aspects, the
inhibitory immune receptor inhibitor specifically binds to a sialic
acid-binding Ig-like lectin (Siglec) receptor (e.g., Siglec-7,
Siglec-9, or both).
[0073] According to certain embodiments, the therapeutic antibody
present in the pharmaceutical composition specifically binds to a
tumor-associated antigen or a tumor-specific antigen. In certain
aspects, the therapeutic antibody present in the pharmaceutical
composition induces antibody-dependent cellular cytotoxicity
(ADCC). In some embodiments, the therapeutic antibody present in
the pharmaceutical composition specifically binds to an antigen
selected from human epidermal growth factor receptor 2 (HER2),
CD19, CD22, CD30, CD33, CD56, CD66/CEACAM5, CD70, CD74, CD79b,
CD138, Nectin-4, Mesothelin, Transmembrane glycoprotein NMB
(GPNMB), Prostate-Specific Membrane Antigen (PSMA), SLC44A4, CA6,
CA-IX, an integrin, C-X-C chemokine receptor type 4 (CXCR4),
cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), and
neuropilin-1 (NRP1). In certain aspects, the therapeutic antibody
present in the pharmaceutical composition is selected from
trastuzumab, cetuximab, daratumumab, girentuximab, panitumumab,
ofatumumab, and rituximab.
[0074] The inhibitory immune receptor inhibitor and therapeutic
antibody can be incorporated into a variety of formulations for
therapeutic administration. More particularly, the inhibitory
immune receptor inhibitor and therapeutic antibody can be
formulated into pharmaceutical compositions by combination with
appropriate, pharmaceutically acceptable excipients or diluents,
and may be formulated into preparations in solid, semi-solid,
liquid or gaseous forms, such as tablets, capsules, powders,
granules, ointments, solutions, injections, inhalants and
aerosols.
[0075] Formulations of the inhibitory immune receptor inhibitor and
therapeutic antibody suitable for administration to the individual
(e.g., suitable for human administration) are generally sterile and
may further be free of detectable pyrogens or other contaminants
contraindicated for administration to a patient according to a
selected route of administration.
[0076] In pharmaceutical dosage forms, the inhibitory immune
receptor inhibitor and therapeutic antibody can be administered in
the form of their pharmaceutically acceptable salts, or they may
also be used alone or in appropriate association, as well as in
combination, with other pharmaceutically active compounds. The
following methods and carriers/excipients are merely examples and
are in no way limiting.
[0077] For oral preparations, the inhibitory immune receptor
inhibitor and therapeutic antibody can be used alone or in
combination with appropriate additives to make tablets, powders,
granules or capsules, for example, with conventional additives,
such as lactose, mannitol, corn starch or potato starch; with
binders, such as crystalline cellulose, cellulose derivatives,
acacia, corn starch or gelatins; with disintegrators, such as corn
starch, potato starch or sodium carboxymethylcellulose; with
lubricants, such as talc or magnesium stearate; and if desired,
with diluents, buffering agents, moistening agents, preservatives
and flavoring agents.
[0078] The inhibitory immune receptor inhibitor and therapeutic
antibody can be formulated for parenteral (e.g., intravenous,
intra-arterial, intraosseous, intramuscular, intracerebral,
intracerebroventricular, intrathecal, subcutaneous, etc.)
administration. In certain aspects, the inhibitory immune receptor
inhibitor and therapeutic antibody is formulated for injection by
dissolving, suspending or emulsifying the conjugate in an aqueous
or non-aqueous solvent, such as vegetable or other similar oils,
synthetic aliphatic acid glycerides, esters of higher aliphatic
acids or propylene glycol; and if desired, with conventional
additives such as solubilizers, isotonic agents, suspending agents,
emulsifying agents, stabilizers and preservatives.
[0079] Pharmaceutical compositions that include the inhibitory
immune receptor inhibitor and therapeutic antibody may be prepared
by mixing the inhibitory immune receptor inhibitor and therapeutic
antibody having the desired degree of purity with optional
physiologically acceptable carriers, excipients, stabilizers,
surfactants, buffers and/or tonicity agents. Acceptable carriers,
excipients and/or stabilizers are nontoxic to recipients at the
dosages and concentrations employed, and include buffers such as
phosphate, citrate, and other organic acids; antioxidants including
ascorbic acid, glutathione, cysteine, methionine and citric acid;
preservatives (such as ethanol, benzyl alcohol, phenol, m-cresol,
p-chlor-m-cresol, methyl or propyl parabens, benzalkonium chloride,
or combinations thereof); amino acids such as arginine, glycine,
omithine, lysine, histidine, glutamic acid, aspartic acid,
isoleucine, leucine, alanine, phenylalanine, tyrosine, tryptophan,
methionine, serine, proline and combinations thereof;
monosaccharides, disaccharides and other carbohydrates; low
molecular weight (less than about 10 residues) polypeptides;
proteins, such as gelatin or serum albumin; chelating agents such
as EDTA; sugars such as trehalose, sucrose, lactose, glucose,
mannose, maltose, galactose, fructose, sorbose, raffinose,
glucosamine, N-methylglucosamine, galactosamine, and neuraminic
acid; and/or non-ionic surfactants such as Tween, Brij Pluronics,
Triton-X, or polyethylene glycol (PEG).
[0080] The pharmaceutical composition may be in a liquid form, a
lyophilized form or a liquid form reconstituted from a lyophilized
form, wherein the lyophilized preparation is to be reconstituted
with a sterile solution prior to administration. The standard
procedure for reconstituting a lyophilized composition is to add
back a volume of pure water (typically equivalent to the volume
removed during lyophilization); however solutions comprising
antibacterial agents may be used for the production of
pharmaceutical compositions for parenteral administration.
[0081] An aqueous formulation of the inhibitory immune receptor
inhibitor and therapeutic antibody may be prepared in a pH-buffered
solution, e.g., at pH ranging from about 4.0 to about 7.0, or from
about 5.0 to about 6.0, or alternatively about 5.5. Examples of
buffers that are suitable for a pH within this range include
phosphate-, histidine-, citrate-, succinate-, acetate-buffers and
other organic acid buffers. The buffer concentration can be from
about 1 mM to about 100 mM, or from about 5 mM to about 50 mM,
depending, e.g., on the buffer and the desired tonicity of the
formulation.
[0082] A tonicity agent may be included in the formulation to
modulate the tonicity of the formulation. Example tonicity agents
include sodium chloride, potassium chloride, glycerin and any
component from the group of amino acids, sugars as well as
combinations thereof. In some embodiments, the aqueous formulation
is isotonic, although hypertonic or hypotonic solutions may be
suitable. The term "isotonic" denotes a solution having the same
tonicity as some other solution with which it is compared, such as
physiological salt solution or serum. Tonicity agents may be used
in an amount of about 5 mM to about 350 mM, e.g., in an amount of
100 mM to 350 mM.
[0083] A surfactant may also be added to the formulation to reduce
aggregation and/or minimize the formation of particulates in the
formulation and/or reduce adsorption. Example surfactants include
polyoxyethylenesorbitan fatty acid esters (Tween), polyoxyethylene
alkyl ethers (Brij), alkylphenylpolyoxyethylene ethers (Triton-X),
polyoxyethylene-polyoxypropylene copolymer (Poloxamer, Pluronic),
and sodium dodecyl sulfate (SDS). Examples of suitable
polyoxyethylenesorbitan-fatty acid esters are polysorbate 20, (sold
under the trademark Tween 20.TM.) and polysorbate 80 (sold under
the trademark Tween 80.TM.). Examples of suitable
polyethylene-polypropylene copolymers are those sold under the
names Pluronic.RTM. F68 or Poloxamer 188.TM.. Examples of suitable
Polyoxyethylene alkyl ethers are those sold under the trademark
Brij.TM.. Example concentrations of surfactant may range from about
0.001% to about 1% w/v.
[0084] A lyoprotectant may also be added in order to protect the
inhibitory immune receptor inhibitor and therapeutic antibody
against destabilizing conditions during a lyophilization process.
For example, known lyoprotectants include sugars (including glucose
and sucrose); polyols (including mannitol, sorbitol and glycerol);
and amino acids (including alanine, glycine and glutamic acid).
Lyoprotectants can be included in an amount of about 10 mM to 500
nM.
[0085] In some embodiments, the pharmaceutical composition includes
the inhibitory immune receptor inhibitor and therapeutic antibody,
and one or more of the above-identified agents (e.g., a surfactant,
a buffer, a stabilizer, a tonicity agent) and is essentially free
of one or more preservatives, such as ethanol, benzyl alcohol,
phenol, m-cresol, p-chlor-m-cresol, methyl or propyl parabens,
benzalkonium chloride, and combinations thereof. In other
embodiments, a preservative is included in the formulation, e.g.,
at concentrations ranging from about 0.001 to about 2% (w/v).
Kits
[0086] As summarized above, the present disclosure provides kits.
According to certain embodiments, the kits include any of the
compositions of the present disclosure. In certain aspects, the
kits include a pharmaceutical composition including an inhibitory
immune receptor inhibitor (e.g., any of the inhibitory immune
receptors described elsewhere herein) and instructions for using
the composition in combination with an antibody therapy being
administered to an individual (e.g., an antibody therapy that
includes administration of a therapeutic antibody that induces
ADCC). The kits of the present disclosure find use, e.g., in
practicing the methods of the present disclosure.
[0087] Kits for practicing the subject methods may include a
quantity of the compositions, present in unit dosages, e.g.,
ampoules, or a multi-dosage format. As such, in certain
embodiments, the kits may include one or more (e.g., two or more)
unit dosages (e.g., ampoules) of a composition that includes an
inhibitory immune receptor inhibitor, or an inhibitory immune
receptor inhibitor and a therapeutic antibody (e.g., a therapeutic
antibody that induces ADCC). In any of the compositions that
include an inhibitory immune receptor inhibitor, the compositions
may include one, two or more inhibitory immune receptor inhibitors,
e.g., two inhibitory immune receptor inhibitors, such as a Siglec-7
inhibitor and a Siglec-9 inhibitor, a Siglec-7 inhibitor and a PD-1
inhibitor, a Siglec-9 inhibitor and a PD-1 inhibitor, or the like.
The term "unit dosage", as used herein, refers to physically
discrete units suitable as unitary dosages for human and animal
subjects, each unit containing a predetermined quantity of the
composition calculated in an amount sufficient to produce the
desired effect. The amount of the unit dosage depends on various
factors, such as the particular inhibitory immune receptor
inhibitor employed, the effect to be achieved, and the
pharmacodynamics associated with the inhibitory immune receptor
inhibitor, the therapeutic antibody, or both, in the subject. In
yet other embodiments, the kits may include a single multi dosage
amount of the composition.
[0088] Components of the kits may be present in separate
containers, or multiple components may be present in a single
container. For example, in a kit that includes both an inhibitory
immune receptor inhibitor and a therapeutic antibody, the
inhibitory immune receptor inhibitor and therapeutic antibody may
be provided in the same composition (e.g., in one or more
containers) or may be provided in separate compositions in separate
containers. Suitable containers include individual tubes (e.g.,
vials), one or more wells of a plate (e.g., a 96-well plate, a
384-well plate, etc.), or the like.
[0089] According to certain embodiments, a kit of the present
disclosure includes instructions for using the compositions to
treat an individual in need thereof. For example, a kit may include
instructions for using the inhibitory immune receptor inhibitor in
combination with a therapeutic antibody that induces ADCC (which
antibody may or may not be present in the kit) to potentiate ADCC
in an individual in need thereof. The instructions may be recorded
on a suitable recording medium. For example, the instructions may
be printed on a substrate, such as paper or plastic, etc. As such,
the instructions may be present in the kits as a package insert, in
the labeling of the container of the kit or components thereof
(i.e., associated with the packaging or sub-packaging) etc. In
other embodiments, the instructions are present as an electronic
storage data file present on a suitable computer readable storage
medium, e.g., portable flash drive, DVD, CD-ROM, diskette, etc. In
yet other embodiments, the actual instructions are not present in
the kit, but means for obtaining the instructions from a remote
source, e.g. via the internet, are provided. An example of this
embodiment is a kit that includes a web address where the
instructions can be viewed and/or from which the instructions can
be downloaded. As with the instructions, the means for obtaining
the instructions is recorded on a suitable substrate.
[0090] The following examples are offered by way of illustration
and not by way of limitation.
EXPERIMENTAL
Example 1--Potentiation of Natural Killer (NK) Cell-Mediated
Antibody-Dependent Cellular Cytotoxicity (ADCC) by Treatment with
Inhibitory Immune Receptor Blocking Antibodies
[0091] Natural Killer cells were purified from human peripheral
blood mononuclear cells isolated from whole blood. They were then
treated with the relevant receptor-blocking antibody (Siglec 7,
Siglec 9 or NKG2D) at 5 microgram/mL for 1 hour at 37.degree. C.
They were then added to BT-20 or Ramos cells at ratios of 4:1
(NK:target cell) along with the therapeutic antibody (trastuzumab
or rituximab) at 10 nM and allowed to incubate for 4 hours. After 4
hours, the cell mixtures were pelleted and the supernatant tested
for levels of lactate dehydrogenase (LDH) released from lysed cells
using a commercial LDH detection kit. From these values were
subtracted measured levels of spontaneously released LDH from NK
and target cells, and then these levels were compared to controls
in which all of the target cells were lysed using a detergent to
give a percent cytotoxicity.
[0092] BT-20 (triple negative breast cancer) cells being treated
with trastuzumab were subjected to the following conditions: (1) no
blocking antibody; (2) Natural Killer Group 2D (NKG2D) blocking
antibody (anti-NKG2D mAb (clone 149810) available from R&D
Systems); (3) Siglec-7 blocking antibody (anti-Siglec-7 mAb (clone
S7.7) available from Biolegend.RTM.); (4) Siglec-9 blocking
antibody (anti-Siglec-9 mAb (clone K8) available from
Biolegend.RTM.); (5) Siglec-7 blocking antibody and Siglec-9
blocking antibody; and (6) incubation with an isotype antibody. As
shown in FIG. 1, the ADCC effect of trastuzamab was potentiated
when the cells were co-treated with Siglec-7 blocking antibody,
Siglec-9 blocking antibody, and a combination of Siglec-7 blocking
antibody and Siglec-9 blocking antibody.
[0093] Ramos (B lymphocyte Burkitt's lymphoma) cells being treated
with rituximab were subjected to the following conditions: (1) no
blocking antibody; (2) Natural Killer Group 2D (NKG2D) blocking
antibody (anti-NKG2D mAb (clone 149810) available from R&D
Systems); (3) Siglec-7 blocking antibody (anti-Siglec-7 mAb (clone
S7.7) available from Biolegend.RTM.); (4) Siglec-9 blocking
antibody (anti-Siglec-9 mAb (clone K8) available from
Biolegend.RTM.); (5) Siglec-7 blocking antibody and Siglec-9
blocking antibody; and (6) incubation with an isotype antibody. As
shown in FIG. 2, the ADCC effect of rituximab was potentiated when
the cells were co-treated with Siglec-7 blocking antibody, Siglec-9
blocking antibody, and a combination of Siglec-7 blocking antibody
and Siglec-9 blocking antibody.
[0094] The data demonstrates a role for inhibitory immune receptors
(in this example, Siglec receptors) in both trastuzumab- and
rituximab-mediated ADCC.
Example 2--Expression of Siglec Ligands
[0095] The expression levels of Siglec ligands on BT-20 and Ramos
cells was determined by flow cytometry. For detection of Siglec
ligands, cells were incubated with a soluble fusion protein that
included the extracellular domain of Siglec-7 or Siglec-9 fused to
a fragment crystallizable (Fc) antibody fragment (Sig-Fc), followed
by incubation with a fluorescent labeled anti-Fc secondary antibody
and detection by flow cytometry. More specifically, Siglec-Fc
fusion proteins were pre-complexed at 5 .mu.g/ml Sig-Fc and 4
.mu.g/ml anti-Fc secondary antibody and incubated with cells for 30
min at 4.degree. C. Cells were then washed 3 times and flow
cytometry was performed. Separately, cells are treated with the
anti-Fc secondary antibody (same 4 .mu.g/ml), then washed as above
and flow cytometry performed.
[0096] Expression levels of Siglec-7 ligands (top) and Siglec-9
ligands (bottom) on BT-20 cells is shown in FIG. 3. Expression
levels of Siglec-7 ligands (top) and Siglec-9 ligands (bottom) on
Ramos cells is shown in FIG. 4.
[0097] The observed increase in fluorescence of the Siglec-Fc
fusion-treated cells over the secondary antibody-only treated cells
indicates that binding was due to the Siglec-Fc fusion protein, and
not the secondary reagent alone. That roughly three orders of
magnitude more signal was observed in the Siglec-Fc fusion-treated
cells indicates that both BT-20 cells and Ramos cells are rich in
Siglec ligands, and provides a basis for why treatment with Siglec
blocking antibodies is effective in potentiating ADCC as
demonstrated in Example 1 above.
Example 3--Siglec-7 Ligand Abundance Predicts Increase in ADCC by a
Trastuzumab-Sialidase Conjugate
[0098] Purified NK cells (isolated as in Example 1), were mixed
with the indicated target cell type at a ratio of 4:1, trastuzumab
was added to a concentration of 10 nM and the cells allowed to
react at 37.degree. C. for 4 hours. In parallel, the same
experiment was setup in which to this mixture of cells was added
sialidase-trastuzumab conjugate to a concentration of 10 nM. At the
end of 4 hours, the cell mixtures were pelleted and percent
cytotoxicity was measured as in Example 1. A `fold increase in
cytotoxicity` was calculated by calculating the quotient of
cytotoxicity of sialidase-pretreated target cells to the
cytotoxicity of untreated cells minus 1. Using flow cytometry and
the labeling technique described in Example 2, the abundance of
both Her2 and cell-surface Siglec-7 ligands was calculated on these
same target cell lines. Ranking cell lines from highest to lowest
Siglec-7 ligand abundance nearly matches that of fold increase in
ADCC cytotoxicity, whereas Her2 levels roughly negatively correlate
with fold increase in cytotoxicity. HER2 expression level is
indicated by number of `+`s. ADCC potentiation is most pronounced
for HER2-low cell line, e.g., as seen in comparing MDA-MB-231 cells
to SKBR3 cells (FIG. 5).
[0099] Notwithstanding the appended claims, the present disclosure
is also defined by the following clauses:
1. A method, comprising:
[0100] administering to an individual receiving an antibody therapy
an inhibitory immune receptor inhibitor.
2. The method according to Clause 1, wherein the inhibitory immune
receptor inhibitor is an antibody or a small molecule. 3. The
method according to Clause 2, wherein the inhibitory immune
receptor inhibitor is an antibody. 4. The method according to
Clause 3, wherein the antibody is selected from the group
consisting of: an IgG, a single chain Fv (scFv), Fab, F(ab').sub.2,
or (scFv').sub.2. 5. The method according to Clause 4, wherein the
antibody is an IgG. 6. The method according to Clause 5, wherein
the IgG is an IgG1, IgG2, IgG3, or IgG4. 7. The method according to
any one of Clauses 3 to 6, wherein the antibody is a monoclonal
antibody. 8. The method according to any one of Clauses 3 to 7,
wherein the antibody is a humanized or human antibody. 9. The
method according to any one of Clauses 1 to 8, wherein the
inhibitory immune receptor inhibitor inhibits an inhibitory immune
receptor present on an immune cell selected from the group
consisting of: a natural killer (NK) cell, a macrophage, a
monocyte, a neutrophil, a dendritic cell, a T cell, a B cell, a
mast cell, a basophil, and an eosinophil. 10. The method according
to any one of Clauses 1 to 9, wherein the inhibitory immune
receptor is a sialic acid-binding Ig-like lectin (Siglec) receptor.
11. The method according to Clause 10, wherein the inhibitory
immune receptor inhibitor inhibits Siglec-7. 12. The method
according to Clause 10, wherein the inhibitory immune receptor
inhibitor inhibits Siglec-9. 13. The method according to any one of
Clauses 1 to 12, wherein the individual has cancer and the method
is for treating the cancer. 14. The method according to Clause 13,
wherein the individual has breast cancer, ovarian cancer, gastric
cancer, colon cancer, renal carcinoma, or a combination thereof.
15. The method according to Clause 13 or Clause 14, wherein the
individual is receiving an antibody therapy that comprises
administering to the individual an antibody that specifically binds
to a tumor-associated antigen. 16. The method according to Clause
13 or Clause 14, wherein the individual is receiving an antibody
therapy that comprises administering to the individual an antibody
that specifically binds to a tumor-specific antigen. 17. The method
according to Clause 13 or Clause 14, wherein the individual is
receiving an antibody therapy that comprises administering to the
individual an antibody that specifically binds an antigen selected
from the group consisting of: human epidermal growth factor
receptor 2 (HER2), CD19, CD22, CD30, CD33, CD56, CD66/CEACAM5,
CD70, CD74, CD79b, CD138, Nectin-4, Mesothelin, Transmembrane
glycoprotein NMB (GPNMB), Prostate-Specific Membrane Antigen
(PSMA), SLC44A4, CA6, CA-IX, an integrin, C-X-C chemokine receptor
type 4 (CXCR4), cytotoxic T-lymphocyte-associated protein 4
(CTLA-4), and neuropilin-1 (NRP1). 18. The method according to any
one of Clauses 13 to 17, wherein the individual is receiving an
antibody therapy that comprises administering to the individual an
antibody that induces antibody-dependent cellular cytotoxicity
(ADCC). 19. The method according to any one of Clauses 1 to 11,
wherein the individual is receiving an antibody therapy that
comprises administering to the individual an antibody selected from
the group consisting of: trastuzumab, cetuximab, daratumumab,
girentuximab, panitumumab, ofatumumab, and rituximab. 20. The
method according to any one of Clauses 1 to 19, comprising
determining the abundance of one or more inhibitory immune receptor
ligands present on cells targeted by the antibody therapy prior to
administering the inhibitory immune receptor inhibitor. 21. The
method according to Clause 20, wherein the inhibitory immune
receptor inhibitor is administered to the individual only upon
determining that the abundance of the one or more inhibitory immune
receptor ligands on cells targeted by the antibody therapy exceeds
a threshold abundance level. 22. The method according to Clause 20
or Clause 21, wherein the abundance of the one or more inhibitory
immune receptor ligands is determined in vivo. 23. The method
according to Clause 22, wherein the abundance of the one or more
inhibitory immune receptor ligands is determined by in vivo
imaging. 24. The method according to Clause 20 or Clause 21,
wherein the abundance of the one or more inhibitory immune receptor
ligands is determined in vitro. 25. The method according to Clause
24, wherein the abundance of the one or more inhibitory immune
receptor ligands is determined on a biopsy sample. 26. The method
according to Clause 25, wherein determining the abundance of one or
more inhibitory immune receptor ligands comprises incubating cells
of the biopsy sample with a reagent comprising an extracellular
domain of the inhibitory immune receptor. 27. The method according
to Clause 26, wherein the reagent comprises a fusion protein
comprising the extracellular domain of the inhibitory immune
receptor. 28. The method according to Clause 27, wherein the fusion
protein comprises a fragment crystallizable (Fc) antibody fragment.
29. The method according to any one of Clauses 20 to 28, wherein
the one or more inhibitory immune receptor ligands comprises a
Siglec-7 ligand. 30. The method according to any one of Clauses 20
to 29, wherein the one or more inhibitory immune receptor ligands
comprises a Siglec-9 ligand. 31. The method according to any one of
Clauses 1 to 30, wherein the inhibitory immune receptor inhibitor
is administered to the individual prior to the onset of the
antibody therapy. 32. The method according to any one of Clauses 1
to 31, wherein the inhibitory immune receptor inhibitor is
administered to the individual concurrently with the antibody
therapy. 33. The method according to Clause 32, wherein the
inhibitory immune receptor inhibitor and the antibody of the
antibody therapy are administered to the individual present in a
same pharmaceutical composition. 34. A pharmaceutical composition,
comprising:
[0101] an inhibitory immune receptor inhibitor;
[0102] a therapeutic antibody; and
[0103] a pharmaceutically acceptable carrier.
35. The pharmaceutical composition of Clause 34, wherein the
inhibitory immune receptor inhibitor is a small molecule. 36. The
pharmaceutical composition of Clause 34, wherein the inhibitory
immune receptor inhibitor is an antibody. 37. The pharmaceutical
composition of Clause 36, wherein the antibody is selected from the
group consisting of: an IgG, a single chain Fv (scFv), Fab,
F(ab').sub.2, or (scFv').sub.2. 38. The pharmaceutical composition
of Clause 36 or Clause 37, wherein the antibody is a monoclonal
antibody. 39. The pharmaceutical composition of any one of Clauses
36 to 38, wherein the antibody is a humanized or human antibody.
40. The pharmaceutical composition of any one of Clauses 34 to 39,
wherein the inhibitory immune receptor inhibitor specifically binds
to an inhibitory immune receptor present on an immune cell selected
from the group consisting of: a natural killer (NK) cell, a
macrophage, a monocyte, a neutrophil, a dendritic cell, a T cell, a
B cell, a mast cell, a basophil, and an eosinophil. 41. The
pharmaceutical composition of any one of Clauses 34 to 40, wherein
the inhibitory immune receptor inhibitor specifically binds to a
sialic acid-binding Ig-like lectin (Siglec) receptor. 42. The
pharmaceutical composition of Clause 41, wherein the inhibitory
immune receptor inhibitor specifically binds to Siglec-7. 43. The
pharmaceutical composition of Clause 41, wherein the inhibitory
immune receptor inhibitor specifically binds to Siglec-9. 44. The
pharmaceutical composition of any one of Clauses 34 to 43,
comprising two or more inhibitory immune receptor inhibitors. 45.
The pharmaceutical composition of Clause 44, comprising two
inhibitory immune receptor inhibitors selected from the group
consisting of: a Siglec-7 inhibitor, a Siglec-9 inhibitor, and a
PD-1 inhibitor. 46. The pharmaceutical composition of any one of
Clauses 34 to 45, wherein the therapeutic antibody specifically
binds to a tumor-associated antigen. 47. The pharmaceutical
composition of any one of Clauses 34 to 45, wherein the therapeutic
antibody specifically binds to a tumor-specific antigen. 48. The
pharmaceutical composition of any one of Clauses 34 to 47, wherein
the therapeutic antibody induces antibody-dependent cellular
cytotoxicity (ADCC). 49. The pharmaceutical composition of any one
of Clauses 34 to 45, wherein the therapeutic antibody specifically
binds to an antigen selected from the group consisting of: human
epidermal growth factor receptor 2 (HER2), CD19, CD22, CD30, CD33,
CD56, CD66/CEACAM5, CD70, CD74, CD79b, CD138, Nectin-4, Mesothelin,
Transmembrane glycoprotein NMB (GPNMB), Prostate-Specific Membrane
Antigen (PSMA), SLC44A4, CA6, CA-IX, an integrin, C-X-C chemokine
receptor type 4 (CXCR4), cytotoxic T-lymphocyte-associated protein
4 (CTLA-4), and neuropilin-1 (NRP1). 50. The pharmaceutical
composition of any one of Clauses 34 to 45, wherein the therapeutic
antibody is selected from the group consisting of: trastuzumab,
cetuximab, daratumumab, girentuximab, panitumumab, ofatumumab, and
rituximab. 51. A kit comprising the pharmaceutical composition of
any one of Clauses 34 to 50. 52. The kit of Clause 51, wherein the
kit comprises the pharmaceutical composition in one or more unit
dosages. 53. The kit of Clause 51 or Clause 52, comprising
instructions for using the composition to treat an individual in
need thereof by antibody-dependent cellular cytotoxicity (ADCC).
54. A kit, comprising: [0104] a pharmaceutical composition
comprising an inhibitory immune receptor inhibitor; and [0105]
instructions for using the composition in combination with an
antibody therapy being administered to an individual. 55. The kit
of Clause 54, wherein the kit comprises the pharmaceutical
composition in one or more unit dosages. 56. The kit of Clause 54
or 55, wherein the inhibitory immune receptor inhibitor is an
antibody. 57. The kit of any one of Clauses 54 to 56, wherein the
inhibitory immune receptor inhibitor inhibits Siglec-7. 58. The kit
of any one of Clauses 54 to 56, wherein the inhibitory immune
receptor inhibitor inhibits Siglec-9. 59. The kit of any one of
Clauses 54 to 58, wherein the pharmaceutical composition comprising
an inhibitory immune receptor inhibitor comprises two or more
inhibitory immune receptor inhibitors. 60. The kit of Clause 59,
wherein the pharmaceutical composition comprising an inhibitory
immune receptor inhibitor comprises two or more inhibitory immune
receptor inhibitors selected from the group consisting of: a
Siglec-7 inhibitor, a Siglec-9 inhibitor, and a PD-1 inhibitor.
[0106] Accordingly, the preceding merely illustrates the principles
of the present disclosure. It will be appreciated that those
skilled in the art will be able to devise various arrangements
which, although not explicitly described or shown herein, embody
the principles of the invention and are included within its spirit
and scope. Furthermore, all examples and conditional language
recited herein are principally intended to aid the reader in
understanding the principles of the invention and the concepts
contributed by the inventors to furthering the art, and are to be
construed as being without limitation to such specifically recited
examples and conditions. Moreover, all statements herein reciting
principles, aspects, and embodiments of the invention as well as
specific examples thereof, are intended to encompass both
structural and functional equivalents thereof. Additionally, it is
intended that such equivalents include both currently known
equivalents and equivalents developed in the future, i.e., any
elements developed that perform the same function, regardless of
structure. The scope of the present invention, therefore, is not
intended to be limited to the exemplary embodiments shown and
described herein. Rather, the scope and spirit of present invention
is embodied by the appended claims.
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